Sponsored,vendor-submitted protocol
Published in
December 2006
2007
(p.13)

Introduction

TissueScan is a unique RT-PCR based system for fast and accurate gene expression profiling across a large number of cancer patient samples. High-quality cDNAs were prepared from 48 well-documented cancer biopsy samples, normalized and assembled into ready-to-use gene expression panels. This alleviates for researchers the tedious work of large sample collection and the meticulous work of RNA/cDNA preparation, while facilitating quick and reliable profiling of gene expression levels across cancer progression stages. TissueScan combines the high sensitivity and specificity of the RT-PCR methodology with a well-designed multi-sample format. It is an excellent tool for validation of potential cancer markers such as those obtained by microarray or differential display.

TissueScan technology was developed to meet the unmet need for quick target validation in cancer biomarker field. TissueScan Oncology products are panels of cDNAs from well-documented biopsy samples. These panels currently represent 4 major types of cancers: lung cancer, colon cancer, ovarian cancer and thyroid cancer. Each sample included in the panels was obtained with the patient's consent and is accompanied by a detailed pathology report. For each cancer panel, 48 tissues were used to produce high-quality cDNA. The samples were designed to represent all progression stages, from stage 0 (pathologically normal appearing tissue) to stage IV. The cDNAs were normalized by beta-actin levels and assembled into a single, 48-well plate. Researchers can simply run a real-time PCR assay by adding PCR master mix and a pair of primers specific for the gene of interest. In mere two hours, you can obtain an accurate expression profile of any gene across 48 tissues of a particular cancer type.

Application example:

To illustrate the validity of TissueScan, we used Lung Cancer Panel (HLRT-101) to evaluate the expression level of Topoisomerase II alpha, which has been known to be upregulated in a large percentage of lung cancers. A pair of primers was designed to detect the transcript of this gene. SYBR Green I based real-time PCR was performed. The raw data (Figure A) and the expression level plot calculated by the Ct method (Figure B) are shown below. A second panel in the kit was used with beta-actin control primers provided in the kit (Figure C).

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